Journal: EMBO Reports

Article Title: Myosin VI regulates ciliogenesis by promoting the turnover of the centrosomal/satellite protein OFD1

doi: 10.15252/embr.202154160

Figure Lengend Snippet: Growth curve of hTERT‐RPE1 cells transfected with myosin VI siRNA. A representative plot of three independent experiments is shown. Growth curve of BJ‐hTERT cells transfected with myosin VI siRNA. A representative plot of two independent experiments is shown. Analysis of DNA content in hTERT‐RPE1 cells stably expressing a myosin VI shRNA. After 10 days of doxycycline induction, the cells were stained with propidium iodide (PI) and analysed by FACS. Ctrl, control cells, not induced. Senescence‐associated β‐gal assay (SA‐β‐gal) of cells treated as in C. Scale bar, 100 μm. IB analysis of hTERT‐RPE1 cells transfected with myosin VI siRNA, with anti‐myosin VI, anti‐p53 and anti‐p21 antibodies. Anti‐GAPDH was used as loading control. Growth curve of hTERT‐RPE1 cells transfected with the indicated p53 and myosin VI siRNAs. A representative plot of three independent experiments is shown. Representative bright‐field images of cells treated with the indicated p53 and myosin VI siRNAs. Scale bar, 200 μm. IB analysis with anti‐myosin VI, anti‐p53 and anti‐p21 antibodies of hTERT‐RPE cells treated with Nutlin‐3, or not treated as control. Anti‐GAPDH was used as loading control. Quantification of OFD1 intensity at the mother or daughter centrioles. hTERT‐RPE1 cells treated or not with Nutlin‐3 were incubated with nocodazole for 1 h (6 μg/ml) and immunostained with anti‐OFD1, anti‐centrin1 and anti‐Cep164 antibodies. Mother centrioles were identified by the coincident staining of centrin1 and Cep164, while daughter centrioles were centrin1‐only stained. Results are expressed as fold change with respect to mock average intensity. Bars represent mean ± SD. Mother centrioles: Mock, n = 148 cells; Nutlin‐3, n = 169 cells, from four independent experiments. Daughter centrioles: Mock, n = 101 cells; Nutlin‐3, n = 122 cells, from three independent experiments. ns, not significant by Mann–Whitney test. Quantification of Cep164 intensity at the mother centrioles in hTERT‐RPE1 cells treated as in (I). Results are expressed as fold change with respect to mock average intensity. Bars represent mean ± SD. Mock, n = 195 cells; Nutlin‐3, n = 196 cells, from four independent experiments. ns, not significant by Mann–Whitney test. Source data are available online for this figure.

Article Snippet: BJ hTERT cells (ATCC) were maintained in DMEM – M199 (4:1), supplemented with 10% FBS and 2 mM l ‐glutamine.

Techniques: Transfection, Stable Transfection, Expressing, shRNA, Staining, Incubation, MANN-WHITNEY

Journal: Cell reports

Article Title: The Toxoplasma Vacuolar H + -ATPase Regulates Intracellular pH and Impacts the Maturation of Essential Secretory Proteins

doi: 10.1016/j.celrep.2019.04.038

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human Foreskin Fibroblast immortalized with hTERT , ATCC , BJ-5ta ATCC® CRL-4001.

Techniques: Recombinant, Modification, Chromatography, Electron Microscopy, Sequencing, Over Expression, Plasmid Preparation, Software